Advantages: No incubation time required.

Measuring Microbial Growth Disadvantages: Cannot always distinguish

between live and dead bacteria. Motile bacteria
Direct Methods of Measurement are difficult to count. Requires a high
1. Plate count: concentration of bacteria (10 million/ml).
Most frequently used method of measuring Indirect Methods of Measurement
bacterial populations. 1. Turbidity:
Inoculate plate with a sample and count number As bacteria multiply in media, it becomes turbid.
of colonies. Use a spectrophotometer to determine %
Assumptions: Each colony originates from a transmission or absorbance.
single bacterial cell. Multiply by a factor to determine concentration.
Original inoculum is homogeneous. Advantages: No incubation time required.
No cell aggregates are present. Disadvantages: Cannot distinguish between
Advantages: Measures viable cells live and dead bacteria. Requires a high
Disadvantages: Takes 24 hours or more for concentration of bacteria (10 to 100 million
visible colonies to appear. Only counts between cells/ml).
25 and 250 colonies are accurate. Must perform 2. Metabolic Activity:
serial dilutions to get appropriate numbers/plate. As bacteria multiply in media, they produce
A. Pour Plate: certain products: Carbon dioxide Acids 4
Introduce a 1.0 or 0.1 ml inoculuminto an Measure metabolic products.
emptyPetri dish. Expensive
Add liquid nutrient medium kept at 50oC. 3. Dry Weight:
Gently mix, allow to solidify, and incubate. Bacteria or fungi in liquid media are centrifuged.
Disadvantages: Not useful for heat sensitive Resulting cell pellet is weighed.
organisms. Colonies appear under agar surface. Doesnt distinguish live and dead cells.
B. Spread Plate:
Introduce a 0.1 ml inoculum onto the surface of
Petri dish.
Spread with a sterile glass rod.
Advantages: Colonies will be on surface and not
exposed to melted agar.
2. Filtration:
Used to measure small quantities of bacteria.
Example: Fecal bacteria in a lake or in ocean
water.
A large sample (100 ml or more) is filtered to
retain bacteria.
Filter is transferred onto a Petri dish.
Incubate and count colonies.
3. Most Probable Number (MPN):
Used mainly to measure bacteria that will not
grow on solid medium.
Dilute a sample repeatedly and inoculate several
broth tubes for each dilution point.
Count the number of positive tubes in each set.
Statistical method: Determines 95% probability
that a bacterial population falls within a certain
range.
4.Direct Microscopic Count:
A specific volume of a bacterial suspension (0.01
ml) is placed on a microscope slide with a
special grid.
Stain is added to visualize bacteria.
Cells are counted and multiplied by a factor to
obtain concentration.